iNOS Antibody (4E5) - BSA Free

Novus Biologicals | Catalog # NBP2-22119

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Flow (Intracellular)

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, In vivo assay, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 4E5

Format

BSA Free
View all iNOS Primary Antibodies »

Product Specifications

Immunogen

iNOS Antibody (4E5) was made to a purified recombinant fragment of human iNOS (C-terminus) expressed in E. coli. [UniProt# P35228]

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

131 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for iNOS Antibody (4E5) - BSA Free

Western Blot: iNOS Antibody (4E5)BSA Free [NBP2-22119]

Western Blot: iNOS Antibody (4E5)BSA Free [NBP2-22119]

Western Blot: iNOS Antibody (4E5) [NBP2-22119] - Analysis using iNOS mouse mAb against Jurkat (1), Jurkat (2), A549 (3), HeLa (4), NIH3T3 (5) and MCF-7 (6) cell lysate.
Immunohistochemistry-Paraffin: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) [NBP2-22119] - Analysis of paraffin-embedded breast cancer tissues using iNOS mouse mAb with DAB staining.
Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) [NBP2-22119] - An intracellular stain was performed on A549 cells with NBP2-22119APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated with antibody at 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).
Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) [NBP2-22119] - An intracellular stain was performed on A549 cells with NBP2-22119PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated with antibody at 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin (PE).
Western Blot: iNOS Antibody (4E5)BSA Free [NBP2-22119]

Western Blot: iNOS Antibody (4E5)BSA Free [NBP2-22119]

Western Blot: iNOS Antibody (4E5) [NBP2-22119] - Analysis using iNOS mAb against human iNOS (AA: 997-1058) recombinant protein. (Expected MW is 32.6 kDa)
Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119] - An intracellular stain was performed on NIH3T3 cells with iNOS Antibody (4E5) NBP2-22119 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).
Immunohistochemistry-Paraffin: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) [NBP2-22119] - Analysis of paraffin-embedded liver cancer tissues using iNOS mouse mAb with DAB staining.
Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119] - An intracellular stain was performed on Caco-2 cells with iNOS Antibody (4E5) NBP2-22119 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).
ELISA: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

ELISA: iNOS Antibody (4E5) - BSA Free [NBP2-22119]

ELISA: iNOS Antibody (4E5) [NBP2-22119] - Red: Control Antigen (100ng); Purple: Antigen (10ng); Green: Antigen (50ng); Blue: Antigen (100ng).

Applications for iNOS Antibody (4E5) - BSA Free

Application
Recommended Usage

ELISA

1:10000

Flow Cytometry

1:200-1:400

Immunohistochemistry

reported in scientific literature (PMID 29891729)

Immunohistochemistry-Frozen

reported in scientific literature (PMID 32780728)

Immunohistochemistry-Paraffin

1:200-1:1000

Western Blot

1:500-1:2000
Application Notes
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 1 review rated 3 using NBP2-22119 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: iNOS

Nitric oxide (NO) is a colorless, free radical gas that carries a variety of messages between cells. Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO. NO production is mediated by members of the nitric oxide synthase (NOS) family including the two constitutive isoforms: brain, bNOS, or neuronal NOS, nNOS (type I) and endothelial cell NOS, eNOS (type III); along with the inducible isoform, iNOS (type II). NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO, requiring the cofactors calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN), heme, and tetrahydrobiopterin (1).

The 131 kDa enzyme, iNOS, is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. While constitutively expressed in kidneys, in other tissues iNOS is induced by bacterial lipopolysaccharides (LPS), growth factors, and cytokines such as IFN-gamma, TNF, IL-1 and IL-2. iNOS is not regulated by the level of intracellular Ca2+ and is constantly active as a dimer when expressed. iNOS activity is elevated in a variety of diseases including atherosclerosis, heart failure, sepsis, solid tumors, and type 2 diabetes. Acting as a critical mediator of inflammation and apoptosis, iNOS inhibitors have been shown to alleviate obesity and stress inducted insulin resistance in mouse models (2,3).

References

1. Forstermann U, and Sessa WC. (2012) Nitric oxide synthases: regulation and function. Eur Heart J. 33(7): 829-837. PMID: 21890489

2. Aktan F. (2004) iNOS-mediated nitric oxide production and its regulation. Life Sci. 75(6):639-53. PMID: 15172174

3. Cinelli MA, Do HT, Miley GP, Silverman RB. (2020) Inducible nitric oxide synthase: Regulation, structure, and inhibition. Med Res Rev. 40(1):158-189. PMID: 31192483

Long Name

Inducible Nitic Oxide Synthase

Alternate Names

NOS2, NOS2A

Entrez Gene IDs

4843 (Human)

Gene Symbol

NOS2

UniProt

P35228

Additional iNOS Products

Product Documents for iNOS Antibody (4E5) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for iNOS Antibody (4E5) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for iNOS Antibody (4E5) - BSA Free

Customer Reviews for iNOS Antibody (4E5) - BSA Free (1)

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  • iNOS Antibody (4E5)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Placental tissue
    Species: Human
    Verified Customer | Posted 01/01/2017
    Non induced, placental tissue on delivery.
    iNOS Antibody (4E5) - BSA Free NBP2-22119

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Protocols

View specific protocols for iNOS Antibody (4E5) - BSA Free (NBP2-22119):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for iNOS Antibody (4E5) - BSA Free

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  • Q: We are having trouble figuring out the correct dilution for IHC. What are the recommended dilutions for this product’s applications?

    A: The dilutions for IHC can vary by catalog number with suggested ranges of anywhere from 1:10 to 1:1000 across our iNOS antibody products. Our recommended dilutions for each application are listed in our products’ data sheets, which can be found on their product webpages. If no dilution guidelines are listed, we recommend running preliminary tests to find the optimal dilution for your experiments.

  • Q: We need to evaluate the level of iNOS in Rat sciatic nerve by Western Blot. I see that you have few antibodies suitable for that, could you please recommend the best what you have for our purposes?

    A: We have 4 iNOS antibodies that are suitable for your experiment. The best choice for you will depend on your specific preferences, in this case the main difference will be the immunogen location. If the immunogen is not important to you, then I would recommend choosing NB300-605 as it gives you the best value and we are able to provide the exact immunogen.

  • Q: We are having trouble figuring out the correct dilution for IHC. What are the recommended dilutions for this product’s applications?

    A: The dilutions for IHC can vary by catalog number with suggested ranges of anywhere from 1:10 to 1:1000 across our iNOS antibody products. Our recommended dilutions for each application are listed in our products’ data sheets, which can be found on their product webpages. If no dilution guidelines are listed, we recommend running preliminary tests to find the optimal dilution for your experiments.

  • Q: We need to evaluate the level of iNOS in Rat sciatic nerve by Western Blot. I see that you have few antibodies suitable for that, could you please recommend the best what you have for our purposes?

    A: We have 4 iNOS antibodies that are suitable for your experiment. The best choice for you will depend on your specific preferences, in this case the main difference will be the immunogen location. If the immunogen is not important to you, then I would recommend choosing NB300-605 as it gives you the best value and we are able to provide the exact immunogen.

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